Molecular biology, which in a broad sense covers the manipulation of DMA and RNA, has become an[unreadable] integral part of most biological research programs, including the UNC Program Project Grant (PPG).[unreadable] Research areas that utilize molecular biology techniques include cloning genes for expression of proteins in[unreadable] cell model systems, site-directed mutagenesis and tagging for cellular localization and functional studies,[unreadable] polymerase chain reaction for cloning and genotyping, RNA isolation and expression analysis, and the[unreadable] development of transgenic mice. The Molecular Biology Core was established at the University of North[unreadable] Carolina (UNC) Cystic Fibrosis Center in 1998 to provide molecular biology services and expertise to faculty[unreadable] who are often primarily trained in cell biology and/or physiology. The Core is currently an integral part of the[unreadable] ongoing PPG to provide primarily cloning and antibody production services. The functions of the Core have[unreadable] been expanded in the latest proposal to suit new project needs, and now include the development and[unreadable] genotyping of transgenic mouse models and RNA expression analyses services. Specifically, the PPG[unreadable] investigators will require the construction of adenoviral, retroviral, and Xenopus expression vectors for[unreadable] expression of a variety of tagged and/or modified proteins, such as CFTR, ENaC subunits, proteases, and[unreadable] nucleotide transporters, in a variety of cell models including well-differentiated human bronchial epithelial[unreadable] (HBE) cell cultures (provided by Core C. Cell Culture). In addition, the Molecular Biology/Mouse Genotyping[unreadable] Core will provide for the generation of new transgenic mouse models (both knock-out and overexpressing[unreadable] models) to study the role of nucleotides and nucleosides release and metabolism in the airway surface liquid[unreadable] regulation and to study the role of proteases in the regulation of ENaC function. Along with this, genotyping[unreadable] of transgenic models for generation of experimental and control animals will also be provided by the[unreadable] Molecular Biology/Mouse Genotyping Core. And finally, RNA isolation and expression analysis services,[unreadable] including access to quantitative real-time PCR equipment and expertise, will be provided. In addition, the[unreadable] Core is continuously striving to provide access to new research tools, including the development of siRNA[unreadable] technologies, and we will seek to make these technologies available for PPG investigators as protocols and[unreadable] strategies are developed. The Molecular Biology/Mouse Genotyping Core will be utilized by all four[unreadable] proposed PPG projects, and thus is an integral component of the efforts outlined in this application.